Apparatus For Producing Tissue Arrays

ABSTRACT

An apparatus for producing a tissue array is described. At least one receiver block ( 1 ), and at least one donor block ( 2 ) that comprises tissue ( 3 ) to be investigated, are provided, a first hollow needle ( 4 ) for creating a cavity in the receiver block ( 1 ), and a second hollow needle ( 5 ) for removing a sample from the tissue ( 3 ) and introducing the sample ( 3 ) into the cavity of the receiver block ( 1 ), being present. A pantograph ( 6 ) is provided for positioning the first and/or the second hollow needle ( 4; 5 ) above the receiver block ( 1 ).

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority of German patent application no. 102005 041 782.5 filed Sep. 1, 2005, which is incorporated by referenceherein. This application also claims priority of German patentapplication no. 10 2005 041 780.9 filed Sep. 1, 2005 and German patentapplication no. 10 2005 041 781.7 filed Sep. 1, 2005, both of which areincorporated by reference herein.

FIELD OF THE INVENTION

The invention concerns an apparatus and method for producing tissuearrays in a paraffin block, such that the block may be sectioned by amicrotome to make a specimen slide having an array of tissue samplesobservable by microscope.

BACKGROUND OF THE INVENTION

Tissue arrays, also called tissue microarrays (TMAs), contain aplurality of different tissue samples in a single receiver block orparaffin block. The receiver block is sectioned in the usual manner witha microtome, and the section is applied onto a specimen slide. Thespecimen slide then contains a plurality of different tissue samples.Because of the large number of tissue samples on a single specimenslide, it is possible to stain or process all the samples under the sameconditions. As a result, even very complex and expensive stainingmethods, for example those derived from immunohistochemistry (IHC) orin-site hybridization (ISH) for revealing DNA or RNA, can be appliedeffectively.

The production of tissue arrays is very time-consuming, however, since aplurality of different samples (up to 1,000) are arranged next to oneanother in one receiver block. From the various tissue or sample blocks,a tissue core is punched out with a hollow needle and transferred into acorrespondingly prepared receiver block.

Before a tissue core is removed from a sample block, the correspondingsite on the sample block must be located and marked. It has provenuseful for this purpose, in practice, first to produce usual microtomesections from a sample block, apply them onto specimen slides, stainthem using a standard method, and have them inspected by a pathologist.The pathologist then selects the sites of interest on the specimenslide, and marks that site directly on the specimen slide.

The laboratory worker producing the tissue arrays then has the task oflocating, on the tissue block or donor block, the sites marked on thespecimen slide, and removing a tissue core at the corresponding sites.

Punched-out portions or paraffin cores are also removed, using a hollowneedle, from the receiver block, which as a rule is made of paraffin.The tissue cores are then introduced into the cavity thus created. Asmentioned, as many as 1,000 tissue cores—depending on theapplication—can be arranged to form an array on one paraffin block. Fromthese dimensions alone, it is apparent that the diameters of the tissuecores are less than 1 mm, and reliable and simple transfer of the tissuecores into the punched hole in the paraffin block is therefore possibleonly with special equipment.

An apparatus for producing a tissue array is known from U.S. Pat. No.6,103,518. This apparatus is characterized in that the receiver block isarranged in stationary fashion, and possesses thereabove a pivotablymounted needle holder for two hollow needles. The needle holder isaligned onto the receiver block by way of an X-Y micrometer displacementdevice. The two needles—one for punching out the receiver block, theother for removing the tissue core—can be brought alternately into theworking position.

For removal of the tissue core from the tissue block, the latter isplaced manually, together with a U-shaped frame, above the paraffinblock and aligned onto the hollow needle.

With this apparatus, it has proven difficult to locate, on the tissueblock, the site marked on the specimen slide, and furthermore to ensurereliable introduction of the tissue core into the paraffin block.

An automated device for producing tissue arrays is known from U.S. Pat.No. 6,383,801 B1. Here both multiple paraffin blocks and multiple tissueblocks are arranged on an X-Y scanning stage. Also provided are twohollow needles operating independently of one another, of which onemakes the punched holes in the paraffin block and the other isresponsible for tissue core removal.

This device is very complex and moreover does not solve the problem ofeasily locating a marked site on the tissue block.

SUMMARY OF THE INVENTION

It is the object of the present invention to improve a manuallyoperating apparatus for producing tissue arrays in such a way thatsimple and reliable introduction of the tissue core into the paraffinblock becomes possible.

This object is achieved, according to the present invention, by thefeatures described in the characterizing portion of claim 1. Furtheradvantageous developments of the invention are the subject matter of thedependent claims.

The invention is characterized in that a pantograph is provided forpositioning the first and/or the second hollow needle above the receiverblock. The pantograph ensures that all movements of the hollow needletake place in one plane, and the tissue core can be reliably positionedabove the receiver block and introduced.

In a further embodiment of the invention, the pantograph comprises twoscissor arms joined to one another, so that a specific conversion ratiocan be implemented by way of different dimensions of the scissor arms.Very accurate positioning of the hollow needles above the tissue blockor the paraffin block can thereby be achieved.

In a development of the invention, the scissor arms are rotatablymounted at their intersection point.

In a further embodiment of the invention, the first scissor armcomprises a receptacle for the first and/or the second hollow needle,and the second scissor arm comprises a receptacle for a detent pin.

In a development of the invention, the detent pin has a positioningarray associated with it, so that positioning of the detent pin on thepositioning array causes that position to be transferred via thepantograph to the hollow needle.

In a further embodiment of the invention, the positioning arraycomprises multiple markings and/or detent holes, and the latter definingthe position of the first and/or the second hollow needle above thereceiver block. It is thereby easily possible to transfer acorresponding map of the positioning array onto the paraffin block. Byexchanging the positioning array for a different array having more orfewer markings or detent holes, different tissue arrays can be producedwithout major modification.

In an advantageous development of the invention, the receiver block isarranged on a turntable so that the receiver block is positionable, viathe turntable, below the hollow needle.

In a development of the invention, the first and the second hollowneedle are arranged next to one another on a needle holder, and theneedle holder is embodied rotatably. The two different needles can thusbe positioned successively above the receiver block, in order on the onehand to make the necessary punched hole in the receiver block and on theother hand to place the tissue core in the punched hole of the receiverblock. The one end of the scissor arm carries a punching lever foractuation of the hollow needle.

In a further embodiment of the invention, in addition to the receiverblock at least one donor block is arranged on the turntable. Thisensures that operations occur in the same plane in terms of the punchedholes or tissue removal, and tissue core placement.

In a development of the invention, multiple first and multiple secondhollow needles, having different diameters, are arranged circularly nextto one another on the needle holder, so that differently dimensionedtissue arrays are also producible. This also ensures that the diameterof the first hollow needle for punching out the receiving block orparaffin block is larger than the diameter of the second hollow needlefor removing the sample.

BRIEF DESCRIPTION OF THE DRAWINGS

The invention will be explained in more detail with reference to anexemplifying embodiment, with the aid of the schematic drawings inwhich:

FIG. 1 is a view of the apparatus having the pantograph; and

FIG. 2 is a detail view of the apparatus having a needle holder.

DETAILED DESCRIPTION OF THE INVENTION

FIG. 1 is a view of the apparatus for producing a tissue array, having areceiver block 1 and a donor block 2 that contains tissue 3 to beinvestigated. Donor block 2 and receiver block 1 are arranged on aturntable 13. Also provided is a pivotably embodied needle holder 14that carries a first hollow needle 4 and a second hollow needle 5. Firsthollow needle 4 serves to create a punched hole in receiver block 1, andsecond hollow needle 5 is provided in order to remove a tissue core fromtissue 3.

The apparatus comprises a pantograph 6, mounted via a rotary shaft 16,having two scissor arms 7 and 8 joined to one another, the end of firstscissor arm 7 carrying a receptacle 9 for a punching lever 15 forselectable actuation of first and second hollow needles 4; 5. The end ofsecond scissor arm 8 is equipped with a detent pin 10 that hasassociated with it a positioning array 11 having individual detent holes12. A positioning knob 19 is located atop detent pin 10.

The position of detent pin 10 on the positioning array is transferredvia the rotatably mounted pantograph to receptacle 9 for punching lever15. Needle holder 14 is joined in positively engaged fashion (FIG. 2) tofirst scissor arm 7, and thereby follows the movement of punching lever15. Selection of the corresponding hollow needle is accomplished by apivoting motion of needle holder 14 about rotary shaft 16. For punching,receiver block 1 is positioned via turntable 13 below the correspondingfirst hollow needle 4; first hollow needle 4 is then driven by punchinglever 15 into receiver block 1 and makes a punched hole therein. Removalof a tissue core from donor block 2, and introduction of the tissue coreinto the punched hole in receiver block 1, are accomplished analogously.Pantograph 6 and positioning array 11 ensure that the positions of firstand second hollow needle 4; 5 are precisely maintained.

It is of course within the scope of the invention to immobilize therespective positions of the turntable by way of additional detents inorder to ensure simple and rapid operation, or also to arrange multipledonor blocks and/or receiver blocks simultaneously.

FIG. 2 is a detail view of needle holder 14, which is mounted rotatablyand displaceably about rotary shaft 16. Needle holder 14 comprises agroove 18 into which a cam 17 positively engages. Cam 17 is joined tofirst scissor arm 7. A movement of scissor arm 7 is transferred via cam17 and groove 18 to needle holder 14.

Precise removal of a tissue core is now described. A fixedly arrangedslider receptacle 22 for a specimen slide holder 20 is provided.Specimen slide holder 20 is arranged movably via a slider 23 in sliderreceptacle 22, and carries a marked specimen slide 25. Present onspecimen slide 25 is a microtome section of donor block 2 having tissue3.

Donor block 2 is arranged below specimen slide holder 20. By way of themovably mounted specimen slide holder 20, congruency is created betweenthe microtome section and tissue 3 in donor block 2.

A sighting device 21 is arranged on second scissor arm 8 via a sightholder 24. Sighting device 21 is aligned onto the marked site onspecimen slide 25 via positioning knob 19. This movement is transferredvia pantograph 6 to punching lever 15 and needle holder 14.

Removal of a tissue core from donor block 2 is then performed by thefact that donor block 2 is rotated via turntable 13 out of its positionbeneath sighting device 21 into a position beneath needle holder 14, andsecond hollow needle 5 is moved via punching lever 15 into tissue 3 inorder to remove a tissue core. Pantograph 6 and sighting device 12ensure that tissue 3 is removed from the site being sighted onto.

PARTS LIST

1 Receiver block, paraffin block

2 Donor block, tissue block

3 Tissue

4 First hollow needle

5 Second hollow needle

6 Pantograph

7 First scissor arm

8 Second scissor arm

9 Receptacle for 4; 5

10 Detent pin

11 Positioning array

12 Detent holes

13 Turntable

14 Needle holder

15 Punching lever

16 Rotary shaft

17 Cam

18 Groove in 14

19 Positioning knob

20 Specimen slide holder

21 Sighting device

22 Slider receptacle

23 Slider

24 Sight holder

25 Specimen slide

1. An apparatus for producing a tissue array comprising: a receiverblock; a donor block including tissue to be investigated; a first hollowneedle for creating a cavity in the receiver block; a second hollowneedle for removing a sample of tissue from the donor block andintroducing the sample into the cavity in the receiver block; and apantograph for positioning at least one of the first hollow needle andthe second hollow needle relative to the receiver block.
 2. Theapparatus for producing a tissue array according to claim 1, wherein thepantograph comprises two scissor arms joined to one another.
 3. Theapparatus for producing a tissue array according to claim 2, wherein thescissor arms are rotatably joined to one another at an intersectionpoint.
 4. The apparatus for producing a tissue array according to claim3, wherein one of the two scissor arms includes a receptacle forreceiving either of the first hollow needle and the second hollowneedle, and another of the two scissor arms includes a detent pin. 5.The apparatus for producing a tissue array according to claim 4, furthercomprising a positioning array, wherein the detent pin is associatedwith the positioning array.
 6. The apparatus for producing a tissuearray according to claim 5, wherein the positioning array comprises aplurality of detent holes for receiving the detent pin, each of theplurality of detent holes defining a position of the receptacle relativeto the receiver block.
 7. The apparatus for producing a tissue arrayaccording to claim 1, further comprising a turntable, wherein thereceiver block is arranged on the turntable.
 8. The apparatus forproducing a tissue array according to claim 7, further comprising: aneedle holder associated with receptacle, wherein the first hollowneedle and the second hollow needle are arranged next to one another onthe needle holder, and the needle holder is movable relative to thereceptacle to align a selected one of the first hollow needle and thesecond hollow needle with the receptacle; and a punching lever alignedwith the receptacle for actuating a hollow needle aligned with thereceptacle.
 9. The apparatus for producing a tissue array according toclaim 7, wherein the donor block is arranged on the turntable.
 10. Theapparatus for producing a tissue array according to claim 8, wherein aplurality of first hollow needles and a plurality of second hollowneedles, having different diameters, are arranged next to one another ina circular arc on the needle holder.
 11. A method of producing a tissuearray comprising the steps of: providing a positioning array having aplurality of predefined positions defined thereon and a pantographoperatively associated with the positioning array; transferring aselected position from the positioning array to a corresponding positionon a receiver block by operating the pantograph; positioning a firsthollow needle at the corresponding position and actuating the firsthollow needle to provide a hole in the receiver block at thecorresponding position; and positioning a second hollow needle carryinga tissue core at the corresponding position and actuating the secondhollow needle to introduce the tissue core into the hole provided in thereceiver block.